WO1998057722A1 - Procede d'elution par gradient - Google Patents

Procede d'elution par gradient Download PDF

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Publication number
WO1998057722A1
WO1998057722A1 PCT/EP1998/003430 EP9803430W WO9857722A1 WO 1998057722 A1 WO1998057722 A1 WO 1998057722A1 EP 9803430 W EP9803430 W EP 9803430W WO 9857722 A1 WO9857722 A1 WO 9857722A1
Authority
WO
WIPO (PCT)
Prior art keywords
elution
separation
gradient
flow rate
isocratic
Prior art date
Application number
PCT/EP1998/003430
Other languages
German (de)
English (en)
Inventor
Karin Cabrera
Alexander Kraus
Edith Dicks
Gerhard Wieland
Dieter Lubda
Original Assignee
Merck Patent Gmbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Merck Patent Gmbh filed Critical Merck Patent Gmbh
Priority to EP98936315A priority Critical patent/EP0989896A1/fr
Priority to JP50368099A priority patent/JP2002504855A/ja
Publication of WO1998057722A1 publication Critical patent/WO1998057722A1/fr

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28042—Shaped bodies; Monolithic structures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
    • B01D15/16—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the fluid carrier
    • B01D15/166—Fluid composition conditioning, e.g. gradient
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
    • B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
    • B01D15/1814—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns recycling of the fraction to be distributed
    • B01D15/1821—Simulated moving beds
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/02—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material
    • B01J20/10—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising silica or silicate
    • B01J20/103—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising silica or silicate comprising silica
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28016—Particle form
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28054—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their surface properties or porosity
    • B01J20/28078—Pore diameter
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
    • B01J20/286—Phases chemically bonded to a substrate, e.g. to silica or to polymers
    • B01J20/287—Non-polar phases; Reversed phases
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
    • B01J20/29—Chiral phases
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3202—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the carrier, support or substrate used for impregnation or coating
    • B01J20/3204—Inorganic carriers, supports or substrates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
    • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
    • B01J20/3244—Non-macromolecular compounds
    • B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2220/00—Aspects relating to sorbent materials
    • B01J2220/50—Aspects relating to the use of sorbent or filter aid materials
    • B01J2220/54—Sorbents specially adapted for analytical or investigative chromatography
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2220/00—Aspects relating to sorbent materials
    • B01J2220/50—Aspects relating to the use of sorbent or filter aid materials
    • B01J2220/58—Use in a single column
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2220/00—Aspects relating to sorbent materials
    • B01J2220/80—Aspects related to sorbents specially adapted for preparative, analytical or investigative chromatography
    • B01J2220/82—Shaped bodies, e.g. monoliths, plugs, tubes, continuous beds
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/28—Control of physical parameters of the fluid carrier
    • G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
    • G01N2030/324—Control of physical parameters of the fluid carrier of pressure or speed speed, flow rate
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/50—Conditioning of the sorbent material or stationary liquid
    • G01N30/52—Physical parameters
    • G01N2030/524—Physical parameters structural properties
    • G01N2030/528—Monolithic sorbent material

Definitions

  • the invention relates to separation methods for liquid chromatography using a new gradient elution method.
  • the method according to the invention shortens the time required for chromatographic separations under isocratic conditions while maintaining the separation performance achieved.
  • the elution behavior is optimized by means of eluent gradients: on the one hand, analytes that are eluted early are to be sufficiently separated, on the other hand, analytes that elute late are not to be eluted with excessive delay so that the separation can be carried out as quickly as possible overall.
  • an eluent gradient means that the sorbent has to be equilibrated again to the starting eluent after each separation. No equilibration is necessary under isocratic conditions, but compromises must be made between the chromatographic separation performance and the time required for the entire separation.
  • the invention relates to methods for separating at least two substances by means of liquid chromatography under isocratic conditions, the flow rate being changed in the course of the elution of the substances to be separated.
  • Figure 1 shows elution diagrams of separations of various vitamins of the vitamin K group: a) separation according to the invention (isocratic, flow gradient chromatography using a monolithic sorbent); b) isocratic using a monolithic sorbent; c) isocratic using a particulate sorbent.
  • the partial representations b) and c) serve for comparison.
  • Experimental details are described in Example 1.
  • Figure 2 shows elution diagrams of separations of the components of a pharmaceutical preparation: a) separation according to the invention (isocratic, flow gradient chromatography using a monolithic sorbent); b) isocratic, at a flow rate of 1 ml / min; c) isocratic, at a flow rate of 3 ml / min d) isocratic, at a flow rate of 5 ml / min.
  • this process is called flow gradient chromatography.
  • sorbents Prerequisites for the use of this method are sorbents in which the H / u curve is sufficiently flat so that the separation performance is practically unchanged over a wide range of the flow rate.
  • sorbents Another prerequisite for the use of flow gradient chromatography is sorbents, in which the flow rate can be varied to a sufficient extent without the operating pressure having to be increased excessively.
  • suitable sorbents are porous ceramic moldings (monolithic sorbents), as are disclosed in WO 94/19687 and in WO 95/03 256.
  • porous ceramic moldings disclosed in WO 95/03 256, which have interconnected macropores and mesopores in the walls of the macropores, the diameter of the macropores having a median value greater than 0.1 ⁇ m, and the diameter of the Mesopores one Median of 2 and 100 nm.
  • Particulate carriers whose pore structure allows the flow rate to be varied without the operating pressure required for this having to be increased excessively should also be applicable to the process according to the invention.
  • the accelerated separation and good separation performance under isocratic conditions which can be achieved by the method according to the invention, are of particular importance for series analysis and for preparative separations.
  • the elution agents customary for isocratic separation processes or for gradient separation processes can be used for the elution process according to the invention.
  • Sorbents for various separation processes such as, for example, reverse phase chromatography, hydrophobic interaction chromatography, ion exchange chromatography or chiral separations, are known to the person skilled in the art. Sorbents derivatized in this way can be used according to the invention. Examples of such separation processes and of suitably derivatized sorbents are disclosed in WO 94/19687.
  • a mixture of vitamins from the K group which are vitamins K1. K2. Contains K3 and K4, is dissolved in acetonitrile water (95: 5; v.v). 10 ⁇ l of this solution are applied to a monolithic column (silica gel, modification RP-18; 83 x 7.2 mm). Then a flow gradient is applied:
  • FIG. 1 b) shows an elution diagram at a constant flow rate (1 ml / min) as above.
  • Figure 1 c) shows the elution diagram using a particulate sorbent (LiChroSpher RP 18; 1 ml / min) as a further comparison.
  • Example 2 Separation of components of a drug
  • a pharmaceutical preparation is dissolved in acetonitrile-20 mM phosphoric acid (90:10; v: v). 5 ⁇ l of this solution are applied to a monolithic column (silica gel, modification RP-18; 93 x 4.6 mm). Then a flow gradient is applied:
  • the elution diagram is shown in Figure 2 a) (detection UV at 254 nm).
  • the order of the eluted components is: maleic acid, paracetamol, caffeine and brompheniramine.
  • Figures 2 b) to 2 d) show the elution diagram at different constant flow rates (sorbent as above):

Landscapes

  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Inorganic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Sustainable Development (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)
  • Solid-Sorbent Or Filter-Aiding Compositions (AREA)

Abstract

L'invention concerne un procédé pour la séparation d'au moins deux substances par chromatographie liquide dans des conditions isocratiques, la vitesse d'écoulement étant modifiée au cours de l'élution (chromatographie à gradients d'écoulement).
PCT/EP1998/003430 1997-06-18 1998-06-08 Procede d'elution par gradient WO1998057722A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP98936315A EP0989896A1 (fr) 1997-06-18 1998-06-08 Procede d'elution par gradient
JP50368099A JP2002504855A (ja) 1997-06-18 1998-06-08 グラディエント溶離法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19725639.2 1997-06-18
DE19725639A DE19725639A1 (de) 1997-06-18 1997-06-18 Gradientenelutionsverfahren

Publications (1)

Publication Number Publication Date
WO1998057722A1 true WO1998057722A1 (fr) 1998-12-23

Family

ID=7832765

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1998/003430 WO1998057722A1 (fr) 1997-06-18 1998-06-08 Procede d'elution par gradient

Country Status (4)

Country Link
EP (1) EP0989896A1 (fr)
JP (1) JP2002504855A (fr)
DE (1) DE19725639A1 (fr)
WO (1) WO1998057722A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004112960A2 (fr) 2003-05-20 2004-12-29 Eksigent Technologies Llc Commande de la vitesse d'ecoulement

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7648761B2 (en) * 2002-10-31 2010-01-19 Merck Patent Gmbh Inorganic monolithic mouldings coated with organic polymers
JP5134636B2 (ja) * 2010-03-01 2013-01-30 株式会社日立ハイテクノロジーズ 高速液体クロマトグラフ装置及び高速液体クロマトグラフ装置の液体送液方法
CN103091413B (zh) * 2012-12-21 2014-10-22 林维宣 一种化妆品中多种增塑剂残留量的检测方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS58171666A (ja) * 1982-03-31 1983-10-08 Shimadzu Corp 液体クロマトグラフ用送液装置
JPH0460456A (ja) * 1990-06-28 1992-02-26 Tosoh Corp 濃度勾配及び／又は流量勾配作製装置
US5236847A (en) * 1990-11-28 1993-08-17 Hitachi, Ltd. Method for analyzing amino acids and apparatus therefor

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS58171666A (ja) * 1982-03-31 1983-10-08 Shimadzu Corp 液体クロマトグラフ用送液装置
JPH0460456A (ja) * 1990-06-28 1992-02-26 Tosoh Corp 濃度勾配及び／又は流量勾配作製装置
US5236847A (en) * 1990-11-28 1993-08-17 Hitachi, Ltd. Method for analyzing amino acids and apparatus therefor

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
KIBBEY T C G ET AL: "High-performance liquid chromatographic analysis of polydisperse ethoxylated non-ionic surfactants in aqueous samples", JOURNAL OF CHROMATOGRAPHY A, vol. 752, no. 1-2, 1 November 1996 (1996-11-01), pages 155-165, XP004071210 *
PATENT ABSTRACTS OF JAPAN vol. 008, no. 010 (P - 248) 18 January 1984 (1984-01-18) *
PATENT ABSTRACTS OF JAPAN vol. 016, no. 252 (P - 1367) 9 June 1992 (1992-06-09) *
SOMMERBURG O ET AL: "Simultaneous detection of carotenoids and vitamin E in human plasma", JOURNAL OF CHROMATOGRAPHY B: BIOMEDICAL SCIENCES & APPLICATIONS, vol. 695, no. 2, 1 August 1997 (1997-08-01), pages 209-215, XP004125785 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004112960A2 (fr) 2003-05-20 2004-12-29 Eksigent Technologies Llc Commande de la vitesse d'ecoulement
EP1631811A2 (fr) * 2003-05-20 2006-03-08 Eksigent Technologie, LLC Commande de la vitesse d'ecoulement
EP1631811A4 (fr) * 2003-05-20 2011-11-02 Eksigent Technologies Llc Commande de la vitesse d'ecoulement
US8673144B2 (en) 2003-05-20 2014-03-18 Dh Technologies Development Pte. Ltd. Flow rate control

Also Published As

Publication number Publication date
JP2002504855A (ja) 2002-02-12
DE19725639A1 (de) 1998-12-24
EP0989896A1 (fr) 2000-04-05

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